Name: Rabbit monoclonal [EPR24334-118] to SARS-CoV-2 nucleocapsid protein
Brand: RabMAb
SKU: ab271180
Rating: 5 (3 reviews)

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Western blot - Anti-SARS-CoV-2 nucleocapsid protein antibody [EPR24334-118] (ab271180)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR24334-118] to SARS-CoV-2 nucleocapsid protein
Suitable for: ELISA, IP, IHC-P, Flow Cyt (Intra), WB, ICC/IF

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free PE

Product name

Rabbit monoclonal [EPR24334-118] to SARS-CoV-2 nucleocapsid protein
See all SARS-CoV-2 nucleocapsid protein primary antibodies
Host species

Rabbit
Tested applications

Suitable for: ELISA, IP, IHC-P, Flow Cyt (Intra), WB, ICC/IFmore details
Species reactivity

Reacts with: SARS-CoV-2
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HEK-293T transfected with myc-His-tagÂ® SARS-CoV-2 Nucleocapsid protein; SARS-CoV-2 Nucleocapsid Recombinant Protein; SARS-CoV-2 infected Vero; ab273530; ab272107 IHC-P: HEK-293T transfected with myc-His-tagÂ® SARS-CoV-2. ICC/IF: 293T transfected with myc-His-tagÂ® SARS-CoV-2; Vero cells after infecting with SARS-CoV-2 virus for 48 h. Flow Cyt (intra): HEK-293T transfected with myc-His-tagÂ® SARS-CoV-2. IP: HEK-293T transfected with myc-His-tagÂ® SARS-CoV-2.
General notes
This antibody is developed with the collaboration of Dr. Hangping Yao, State Key Laboratory for Diagnosis and Treatment of Infectious Diseases in Zhejiang University, School of Medicine.

This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR24334-118
Isotype

IgG
Research areas

Alternative Versions
- Anti-SARS-CoV-2 nucleocapsid protein antibody [EPR24334-118] - BSA and Azide free (ab280201)
Compatible Secondaries
Dyes/Markers
- Alexa Fluor® 488 Anti-Myc tag antibody [9E10] (ab202008)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab271180 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
ELISA		Use at an assay dependent concentration. Use at 31.25 ng /ml
IP		1/30.
IHC-P		1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt (Intra)		1/500.
WB	(2)	1/1000.
ICC/IF	(1)	1/1000.

Notes
ELISA Use at an assay dependent concentration. Use at 31.25 ng /ml
IP 1/30.
IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt (Intra) 1/500.
WB 1/1000.
ICC/IF 1/1000.

Relevance

Nucleocapsid protein is a most abundant protein of coronavirus on the helical nucleocapsid of coronaviruses. N protein of SARS CoV-2 ab273530 is a structural protein required for RNA synthesis, and has RNA chaperone activity that may be involved in template switch. N protein enters the host cell with the viral RNA to facilitate its replication and process the virus particle assembly and release. N protein is a highly immunogenic phosphoprotein also implicated in modulating cell signalling pathways. Coronavirus nucleocapsid proteins localize to the cytoplasm and the nucleolus, a subnuclear structure, in both virus-infected primary cells and in cells transfected with plasmids that express N protein.
Alternative names
- 2019-nCoV nucleocapsid protein antibody
- N antibody
- NC antibody
- Nucleoprotein antibody
- Protein N antibody
see all

Western blot - Anti-SARS-CoV-2 nucleocapsid protein antibody [EPR24334-118] (ab271180)

All lanes : Rabbit monoclonal [EPR24334-118] to SARS-CoV-2 nucleocapsid protein (ab271180) at 1/1000 dilution

Lane 1 : HEK-293T (human embryonic kidney) transfected with SARS-CoV-2 Nucleocapsid protein expression vector containing a myc-His-tag®, whole cell lysate
Lane 2 : HEK-293T (human embryonic kidney) transfected with SARS-CoV-1 Nucleocapsid protein expression vector containing a myc-His-tag®, whole cell lysate
Lane 3 : HEK-293T (human embryonic kidney) transfected with MERS Nucleocapsid protein expression vector containing a myc-His-tag®, whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Observed band size: 51 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Exposure time 7.75 seconds
Western blot - Anti-SARS-CoV-2 nucleocapsid protein antibody [EPR24334-118] (ab271180)

All lanes : Rabbit monoclonal [EPR24334-118] to SARS-CoV-2 nucleocapsid protein (ab271180) at 1/1000 dilution

Lane 1 : SARS-CoV-2 Nucleocapsid Recombinant Protein at 0.1 µg
Lane 2 : SARS-CoV-2 infected Vero for 48 hours
Lane 3 : Uninfected Vero

Observed band size: 51 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Data was kindly provided by Dr. Hangping Yao, State Key Laboratory for Diagnosis and Treatment of Infectious Diseases in Zhejiang University, School of Medicine.
Western blot - Anti-SARS-CoV-2 nucleocapsid protein antibody [EPR24334-118] (ab271180)

All lanes : Rabbit monoclonal [EPR24334-118] to SARS-CoV-2 nucleocapsid protein (ab271180) at 1/1000 dilution

Lanes 1 & 4 : Recombinant Human coronavirus SARS-CoV-2 nucleocapsid protein (His tag) (ab273530)
Lanes 2 & 5 : Recombinant Human coronavirus SARS Nucleocapsid Protein (ab270829)
Lanes 3 & 6 : Recombinant Human coronavirus SARS-CoV-2 Nucleocapsid Protein (His tag) (ab272107)

Lysates/proteins at 0.1 µg per lane.

Performed under reducing conditions.

Observed band size: 51 kDa why is the actual band size different from the predicted?

Lanes 1 - 3: Green - ab271180 observed at 51 kDa. Lanes 4 - 6: Red - loading control ab18184 (Mouse monoclonal [HIS.H8] to 6X His tag^®) observed at 51 kDa.

ab271180 was shown to react with SARS-CoV-2 nucleocapsid protein in Western blot. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab271180 and ab18184 (Mouse monoclonal [HIS.H8] to 6X His tag^®) overnight at 4 °C at a 1 in 1000 dilution of both antibodies. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
Western blot - Anti-SARS-CoV-2 nucleocapsid protein antibody [EPR24334-118] (ab271180)

Samples

Lanes 1 & 3: Recombinant Human coronavirus SARS-CoV-2 Nucleocapsid Protein (His tag) (ab273530) (0.2 µg)

Lanes 2 & 4: Recombinant Human coronavirus SARS-CoV-2 Nucleocapsid Protein (His tag) (ab272107) (0.2 µg)

Green: Rabbit monoclonal [EPR24334-118] to SARS-CoV-2 nucleocapsid protein (ab271180)

Red: Mouse monoclonal [HIS.H8] to 6X His tag® (ab18184)

Lanes 1 & 2: Green - ab271180 observed at 50 kDa.

Lanes 3 & 4: Red - loading control Mouse anti-6x HisTag observed at 50 kDa.

ab271180 was shown to react with SARS-CoV-2 nucleocapsid protein in Western blot. Membranes were blocked in 3% milk before incubation with ab271180 and Mouse anti-6x HisTag (ab18184) overnight at 4 °C at a 1/1000 dilution. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1/20000 dilution for 1 h at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SARS-CoV-2 nucleocapsid protein antibody [EPR24334-118] (ab271180)

Immunohistochemical analysis of paraffin-embedded Human lung labeling SARS-CoV-2 with ab271180 at 1/4000 dilution followed by ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Negative control: No staining on human lung. The section was incubated with ab271180 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SARS-CoV-2 nucleocapsid protein antibody [EPR24334-118] (ab271180)

Immunohistochemical analysis of paraffin-embedded HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with SARS-CoV-2 Nucleocapsid protein expression vector containing a myc-His-tag® (Panel A)
HEK-293T transfected with SARS-CoV-1 Nucleocapsid protein expression vector containing a myc-His-tag® (Panel B) HEK-293T transfected with MERS Nucleocapsid protein expression vector containing a myc-His-tag® (Panel C) HEK-293T transfected with control vector containing a myc-His-tag® (Panel D) with 271180 at 1/4000 dilution followed by ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on (A) HEK-293T transfected with a SARS-CoV-2 Nucleocapsid protein expression vector is observed. No staining on (B) HEK-293T transfected with a SARS-CoV-1 Nucleocapsid protein expression vector, (C) HEK-293T transfected with MERS Nucleocapsid protein expression vector and (D) HEK-293T transfected with empty vector. The section was incubated with ab271180 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SARS-CoV-2 nucleocapsid protein antibody [EPR24334-118] (ab271180)

Immunohistochemical analysis of paraffin-embedded Mouse lung labeling SARS-CoV-2 with ab271180 at 1/4000 dilution followed by ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Negative control: No staining on Mouse lung. The section was incubated with ab271180 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SARS-CoV-2 nucleocapsid protein antibody [EPR24334-118] (ab271180)

Immunohistochemical analysis of paraffin-embedded Rat lung labeling SARS-CoV-2 with ab271180 at 1/4000 dilution followed by ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Negative control: No staining on Rat lung. The section was incubated with ab271180 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunocytochemistry/ Immunofluorescence - Rabbit monoclonal [EPR24334-118] to SARS-CoV-2 nucleocapsid protein (ab271180)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100-permeabilized 293T cells transfected with myc-tagged SARS-CoV-2 Nucleocapsid protein expression vector labelling SARS-CoV-2 with ab271180 at 1/1000 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution (Red). ab202008 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 488) was used to counterstain Myc tag at 1/200 dilution (Green). The nuclear counterstain was DAPI (Blue). Confocal image showing cytoplasmic staining in 293T cells transfected with myc-tagged SARS-CoV-2 Nucleocapsid protein expression vector.
Immunocytochemistry/ Immunofluorescence - Rabbit monoclonal [EPR24334-118] to SARS-CoV-2 nucleocapsid protein (ab271180)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100-permeabilized Vero cells labelling SARS-CoV-2 with ab271180 at 1/1000 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/500 dilution. Fluorescent image showing positve staining in Vero cells after infecting with SARS-CoV-2 virus for 48 h. Nuclear counter stain DAPI.
Two indenpendent optical fields were captured for imaging at each given antibody dilution.

This piece of data was kindly provided by Dr. Hangping Yao, State Key Laboratory for Diagnosis and Treatment of Infectious Diseases in Zhejiang University, School of Medicine
Immunocytochemistry/ Immunofluorescence - Rabbit monoclonal [EPR24334-118] to SARS-CoV-2 nucleocapsid protein (ab271180)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100-permeabilized 293T cells transfected with myc-tagged SARS-CoV-1 and MERS Nucleocapsid protein labelling SARS-CoV-2 with ab271180 at 1/1000 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution. ab202008 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 488) was used to counterstain Myc tag at 1/200 dilution (Green). The nuclear counterstain was DAPI (Blue). Confocal image showing no staining in 293T cells transfected with myc-tagged SARS-CoV-1 and MERS Nucleocapsid protein, which are negative controls for SARS-CoV-2.
Flow Cytometry (Intracellular) - Anti-SARS-CoV-2 nucleocapsid protein antibody [EPR24334-118] (ab271180)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HEK-293T (human embryonic kidney) transfected with myc-tagged SARS-CoV-2 Nucleocapsid protein expression construct labelling SARS-CoV-2 with ab271180 at 1/500 dilution (Right) compared with a Rabbit monoclonal IgG (ab172730) (Left) isotype control.

A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077 at 1/2000 dilution was used as the secondary antibody.
Flow Cytometry (Intracellular) - Anti-SARS-CoV-2 nucleocapsid protein antibody [EPR24334-118] (ab271180)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HEK-293T (human embryonic kidney) transfected with myc-tagged SARS-CoV-1 Nucleocapsid protein expression construct (Left)/ HEK-293T (human embryonic kidney) transfected with myc-tagged MERS Nucleocapsid protein expression construct (Right).

A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077 at 1/2000 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-SARS-CoV-2 nucleocapsid protein antibody [EPR24334-118] (ab271180)

SARS-CoV-2 was immunoprecipitated from 0.35 mg HEK-293T (human embryonic kidney) transfected with SARS-CoV-2 Nucleocapsid protein expression vector containing a myc-His-tag®, whole cell lysate 5 µg with ab271180 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab271180 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: HEK-293T (human embryonic kidney) transfected with SARS-CoV-2 Nucleocapsid protein expression vector containing a myc-His-tag® whole cell lysate 5 μg

Lane 2: ab271180 IP in HEK-293T transfected with SARS-CoV-2 Nucleocapsid protein expression vector containing a myc-His-tag® whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab271180 in HEK-293T transfected with SARS-CoV-2 Nucleocapsid protein expression vector containing a myc-His-tag® whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3.25 seconds.
ELISA - Anti-SARS-CoV-2 nucleocapsid protein antibody [EPR24334-118] (ab271180)

Plates were coated with Recombinant Human coronavirus SARS-CoV-2 nucleocapsid protein (His tag, ab273530) and Recombinant Human coronavirus SARS Nucleocapsid Protein (ab270829) at 500 ng/ml.

Primary antibody (ab271180) concentration range 0.045-100 ng /ml.

Pre-adsorbed secondary antibody, goat anti-rabbit IgG H&L (HRP, ab97080) used at 1/2000 dilution.

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

SDS download

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Datasheet download

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Publishing research using ab271180? Please let us know so that we can cite the reference in this datasheet.

ab271180 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Application

Immunocytochemistry/ Immunofluorescence

Sample

SARS-CoV-2 Cell (human ACE-2 expressing A549 cells)

Permeabilization

Yes - BD Permwash

Specification

human ACE-2 expressing A549 cells

Blocking step

BD Permwash as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 37°C

Fixative

10% formalin

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Tessa Prince

Verified customer

Submitted May 13 2021

Application

Western blot

Sample

African green monkey Cell lysate - whole cell (Vero E6)

Gel Running Conditions

Reduced Denaturing (NuPAGE Bis-Tris 4-12%)

Loading amount

5 µg

Treatment

SARS CoV2 variants 24hpi

Specification

Vero E6

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Stuart Armstrong

Verified customer

Submitted May 12 2021

Application

Western blot

Sample

Human Cell lysate - whole cell (Caco-2 human colorectal adenocarcinoma cells)

Gel Running Conditions

Reduced Denaturing (4-12% Bis-Tris NuPAGE)

Loading amount

40 µg

Specification

Caco-2 human colorectal adenocarcinoma cells

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MS. Nicola Goodman

Verified customer

Submitted Apr 30 2021

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Key features and details

Related conjugates and formulations

Overview

Product name

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Dyes/Markers

Isotype control

Applications

The Abpromise guarantee

Target

Relevance

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (0)

Customer reviews and Q&As

Immunocytochemistry/ Immunofluorescence abreview for Anti-SARS-CoV-2 nucleocapsid protein antibody [EPR24334-118]

Western blot abreview for Anti-SARS-CoV-2 nucleocapsid protein antibody [EPR24334-118]

Western blot abreview for Anti-SARS-CoV-2 nucleocapsid protein antibody [EPR24334-118]